五味子乙素对染矽尘大鼠肺组织转化生长因子β1信号转导通路分子mRNA表达的影响

目的 观察五味子乙素(sehisandrin B,Sch-B)对染矽尘大鼠肺组织转化生长因子β1(TGF-β1)信号通路分子mRNA表达的影响,探讨其防治矽肺纤维化的作用及机制.方法 将96只Wistar大鼠随机分为对照组、染矽尘组、Sch-B干预组,每组32只,气管暴露法一次性染尘建立大鼠矽肺模型,染矽尘组和Sch-B干预组每只大鼠气管内注入1 ml(50 mg/ml)SiO2混悬液,对照组注入1 ml灭菌生理盐水.染尘后第1天开始灌胃干预,Sch-B组给予Sch-B(80mg·kg-1/d-1),染矽尘组和对照组给予等体积的橄榄油.不同处理3、7、14和28 d后,各组处死8只大鼠,对肺组织HE染色观察病理改变;用反转录-聚合酶链式反应(RT-PCR)法测定大鼠肺组织中TGF-β1、转化生长因子βⅡ型受体(TGF-βRⅡ)和Smad4基因表达.结果 染矽尘组肺损伤明显,在3和7 d时表现为明显的肺泡炎,可见肺泡间隔水肿,大量炎性细胞浸润,在14d时,肺泡间隔明显增宽,成纤维细胞和胶原基质明显增多;在28 d时,肺泡结构破坏,肺泡壁明显增厚,肺组织以胶原沉积和肺纤维化改变为主.Sch-B组肺泡炎和肺纤维化程度均较染矽尘组明显减轻.染矽尘组3、7、14、28 d时大鼠肺内TGF-β1、TGF-βRⅡ和Smad4的mRNA表达(TGF-β1:1.03±0.31、1.33±0.39、1.08±0.26、0.82±0.16,TGF-βRⅡ:0.65±0.11、0.80±0.16、0.83±0.24、0.62±0.15,Smad4:0.87±0.15、0.68±0.11、0.78±0.19、0.30±0.08)均明显高于对照组(TGF-β1:0.59±0.22、0.55±0.25、0.56±0.20、0.55±0.12,TGR-βRⅡ:0.28±0.13、0.31 ±0.15、0.34±0.15、0.27±0.09,Smad4:0.23±0.11、0.40±0.12、0.39±0.12、0.18±0.06),差异均有统计学意义(P＜0.01或P＜0.05),其中TGF-β1在第7天时达高峰后开始下降.Sch-B干预组各时间点TGF-β1和Smad4(TGF-β1:0.68±0.28、0.88±0.25、0.75±0.11、0.61±0.14,Smad4:0.25±0.12、0.45±0.09、0.44±0.07、0.21±0.04)均低于染矽尘组,差异有统计学意义(P＜0.01或P＜0.05).与染矽尘组相比,Sch-B干预组TGF-βRⅡmRNA表达无明显改变,差异无统计学意义(P＞0.05).结论 Sch-B能减轻染矽尘大鼠肺组织的纤维化程度,其作用机制可能是通过抑制TGF-β1和Smad4的mRNA表达上调,从而干扰TGF-β1/Smad4信号通路对靶基因的激活来实现的.

Abstract：

Objective To investigate the effects of schisandrin B (Sch-B) on expression of transforming growth factor-β1 (TGF-β1) and signal transduction molecule mRNA in rat lungs exposed to SiO2,and explore the intervention mechanism of Sch-B on pulmonary fibrosis induced by SiO2. Methods Ninety six Wistar rats were randomly divided into control (normal saline) group, SiO2 group and SiO2 plus Sch-B group.The rats were exposed to SiO2 by direct tracheal instillation to establish the silicotic animal models. SiO2 group and SiO2 plus Sch-B group were treated with 1ml SiO2 (50 mg/ml) for each rat From the first day after model establishment, SiO2 plus Sch-B group were orally given Sch-B (80 mg/kg) a day, control group and silica group were orally given olive oil. On the 3rd, 7th,14th and 28th days after treatment, 8 rats in each group were sacrificed and samples were collected. The histo-pathological examination of lung was performed by HE staining. The expression levels of TGF-β1 、TGFβR Ⅱ and Smad4 mRNA in the lung tissues were detected by RT-PCR. Results The results of histo-pathological examination showed that in SiO2 group, lung tissues were injured obviously; the alveolar inflammation with alveolus interval edema and inflammation cell infiltration appeared on the 3rd and 7th days; the alveolus interval became thicker, became thicker, fibroblast and collagen matiix increased markedly on 14th day; the alveolar structure was damaged, alveolar wall thickened obviously,collagen aggradation and pulmonary fibrosis displayed on 28th day. The alveolar inflammation and pulmonary fibrosis in SiO2 plus Sch-B group were significantly less than those in SiO2 group. The expressions levels of TGF-β1、TGFβR Ⅱ and Smad4 mRNA (TGF-β1: 1.03±0.31 、1.33±0.39、1.08±0.26、0.82±0.16,TGF-βR Ⅱ:0.65 ±0.11、0.80 ±0.16、0.83 ±0.24、0.62 ±0.15, Smad4: 0.87 ±0. 15、0.68 ±0.11、0.78 ±0.19、0.30 ±0.08) in SiO2group were significantly higher than those in the control group (TGF-β1:0.59±0.22、0.55 ±0.25、0.56±0.20、0.55 ±0.12,TGR-βR Ⅱ:0.28 ±0.13、0.31 ±0. 15、0.34 ±0.15、0.27 ±0.09,Smad4:0.23 ±0.11、0.40 ±0. 12、0.39 ±0.12、0.18±0.06)(P＜0.01 or P＜0.05), but the expression level of TGF-β1 mRNA was the highest on the 7th day. The expression levels of TGF-β1 and Smad4 mRNA (TGF-β1: 0.68 ±0.28、0.88 ±0.25、0.75 ±0.11、0.61 ±0. 14,Smad4:0.25 ±0.12、0.45 ±0.09、0.44 ±0.07、0.21 ±0.04) in SiO2 plus Sch-B group were significantly lower than those in SiO2 group (P＜0.01 or P＜0.05),but there were no significant differences of the TGFβR Ⅱ mRNA expression levels between SiO2 group and SiO2 plus Sch-B group. Conclusion Sch-B can reduce the pulmonary fibrosis induced by SiO2 through inhibition of the mRNA express of TGF-β1 and Smad4 in the lung tissue, modulating the TGF-β1/Smad4 signal transduction pathway and inhibiting the target gene activation.     

Effects of silicotic alveolar macrophages exposed to SiO2 on the expression of type Ⅲ collagen and type Ⅲ procollagen in human lung fibroblasts

Objective To study the effects of supernatant of alveolar macrophages (AM) exposed to SiO2 on the expression of type Ⅲ collagen and type Ⅲ procollagen in human lung fibroblasts (HELF) and the intervention effects of anti-TGF-β1 antibody Methods AMs collected from a silicotic by bronchoalveolar lavage were divided into 2 parts, one part was exposed to SiO2 and other part served as control. The supernatant was obtained from AMs cultured for 18 h. HELF were divided into (1) exposure group, which was added with supernatant from AMs exposed to SiO2; (2) control group, which was added with the supernatant from AMs not exposed to SiO2; (3) blank control group, which was added with DMEM; (4) exposure group plus anti-TGF-β1antibody (10μg/ml); (5) control group plus anti-TGF-β1 antibody (10 μg/ml). (1)-(3) groups were cultured for 6, 12, 18, 24, 36, 48h, respectively. (4)-(5) groups were cultured for 18, 24, 36, respectively.Immunocytochemical test and Western blot assay were used to detect pC Ⅲ expression levels in HELF and C Ⅲ expression levels in the supernatant of HELF culture, respectively. Results The pC Ⅲ expression levels of exposure group were 0.1423 ±0.0107,0.1624±0.0011,0.1925 ±0.0050,0.2421 ±0.0097 and 0.2103 ±0.0103,respectively, which were significantly higher than those (0.1212±0.0079、0.1414±0.0058、0.1620±0.0081、0.1965±0.0103 、0.1715±0.0116) of control group (P＜0.05 or P＜0.01). The C Ⅲ levels of exposure group were (0.2559±0.0061、0.3249±0.0110、0.4171±0.0193、0.5441 ±0.0452、0.4751±0.0252), respectively, which were significantly higher than control group (0.2296±0.0121、0.2778±0.0116、0.3367±0.0269、0.3722±0.0214). The pC Ⅲ and C Ⅲ expression levels of exposure plus anti-TGF-β1 antibody group were significantly lower than those of control plus anti-TGF-β1 antibody group (P＜0.05 or P＜0.01).Conclusion AMs exposed to SiO2 can induce the elevated pC Ⅲ and C Ⅲ expression levels in HELF by TGF-β 1 to some extent.     

姜黄素对大鼠肺间质纤维化影响的机制研究

Objective To observe the possible mechanism and inhibitory effects of curcumin on pulmonary fibrosis induced bleomycin in rats at the fibrosing stage. Methods 80 male Sprague-Dawley rats were random divided into 4 groups (20 rats in each group). Rats in the fibrosis model group, the prednisone group and the curcumin group were induced by instilled bleomycin through tracheal, rats in the control group with same volume normal saline. Since the 15th day after bleomycin administration, the curcumin group and prednisone group were given curcumin (300 mg/kg) or prednisone (5mg/kg) per day by intragastric administration, respectively. The normal control group and the model group were given 1% sodium carboxymethyl cellulose ( 10ml/kg). Six rats of each group were random sacrificed on the 21st, 28th, 42nd and 56th days after bleomycin administration. The histological changes of the pulmonary were evaluated by H. E and Masson dyeing. The expressions of transforming growth factor-β1 (TGF-β1), platelet-derived growth factor (PDGF) and hydroxyproline in the tissue of pulmonary were assessed by immunohistochemistry and digestion method. Results Pulmonary fibrosis and hydroxyproline level in the curcumin group were obviously reduced as compared with the model group on the 42nd and 56th day[42 d:1. 28 ±0. 61 vs 2. 28 ±0. 39,P <0. 01 ;(1.73 ±0. 22)mg/g vs (2.50 ±0. 37) mg/g, P <0.01;56 d:1.00 ±0.59 vs 1.73 ±0.36, P< 0. 05; ( 1.57 ± 0. 36) mg/g vs (2. 20 ± 0. 42) mg/g, P < 0. 01 ], and it was also lower than that in prednisone group on the 42nd day( P < 0. 05 ). The expression of TGF-β1 and PDGF in the curcumin group were obviously lower than that in the model group on the 28th, 42nd and 56th day[28 d:TGF-β1 :3642. 05 ±839. 31 vs 5067. 35 ±738. 39, P <0. 05 ;PDGF:2957. 55 ±739. 16 vs 4457. 75 ±568. 39, P <0. 05;42 d: TGF-β1: 2689. 73 ± 529.22 vs 4089. 50 ± 619. 37, P < 0. 01; PDGF: 2834. 46 ± 567. 16 vs 3239. 52 ±628. 26, P <0. 01 ;56 d:TGF-β1: 1968.57 ±408. 36 vs 2968.20 ±498.42, P <0. 01 ;PDGF: 1083.36 ±381.35 vs 2019. 40 ±412. 36, P <0. 01 ], which was lower than that in prednisone group on the 42nd and 56th day (42 d,TGF-β1 :3529. 07 ±981.35,PDGF:2618. 34 ±813. 34;56 d,TGF-β1 :2530. 83 ±439. 37,PDGF: 1738. 35 ±536. 62, Pall <0. 05 ) , and it had no obvious difference compared with control group on the 56th day ( P > 0. 05 ). Conclusion Curcumin could alleviate bleomycin-induced pulmonary fibrosis in rats at the fibrosing stage by inhibiting the expressions of TGF-β1 and PDGF.     

石英尘和博莱霉素诱导的大鼠肺纤维化模型的比较研究

目的 比较石英尘和博莱霉素诱导的大鼠肺纤维化模型中的肺泡炎和早期纤维化改变,并对其机制进行探讨.方法 将大鼠随机分为SiO2组(14只,气管内注入40mg/ml SiO2混悬液1 ml)、BLM组(14只,气管内注入5 mg/kg BLM A5)和对照组(14只,气管内注入1 ml无菌生理盐水).在造模后7、14 d每组各处死7只动物,取肺组织病理切片行HE染色,对肺泡炎进行计分,行饱和苦味酸天狼猩红染色,采用图像分析系统测得各组胶原面积后,进行定量分析,免疫组织化学技术检测肺组织中CD68和TNF-α蛋白表达情况,采用图像分析计算累积吸光度值,进行半定量分析.结果 (1)HE染色光学显微镜下可见,BLM组7d时肺泡炎(肺泡炎评分2.814±0.832)最明显,偏振光下显示BLM组14d时肺纤维化[胶原面积(1284.57±554.72)μm2)]最严重,均明显高于对照组和同期SiO2组,差异有统计学意义(P＜0,05).(2)免疫组化结果显示,BLM组7 d时TNF-α表达最高(17.100±1.831),明显高于对照组(0.420±0.020)、SiO2组7 d(7.909±1.275)及BLM组14d(13.506±1.454),差异均有统计学意义(P＜0.05);在SiO2组14 d时TNF-α表达为22.778±2.512,明显高于BLM组(14 d)、对照组及SiO2组(7 d),差异有统计学意义(P＜0.05).14 d时,SiO2组CD68表达明显高于对照组、BLM组(14 d)及SiO2组(7d),差异有统计学意义(P＜0.05).结论 BLM诱导的大鼠肺损伤模型的早期肺泡炎重于SiO2诱导的大鼠肺损伤模型,纤维化进程早于SiO2诱导的大鼠肺损伤模型,TNF-α在两种模型的病程中均起着重要的作用,而巨噬细胞更为持续地参与了SiO2诱导的肺纤维化.

Abstract：

Objective To compare the pulmonary alveolitis and the early fibrosis of pulmonary fibrosis induced by quartz dust and bleomycin in rats, and investigate their mechanism. Methods The female rats were divided into three groups: control group exposed to normal saline by the trachea; SiO2 group exposed to SiO2 by the trachea; BLM group exposed to BLM A5 by the trachea. Each half of the animals were sacrificed on the 7th andl4th day after expoasure. The lungs of rats were collected to observe pulmonary alveolitis by HE staining and to observe fibrosis by saturated picric acid sirius red staining. The expression of tumor necrosis factor-α (TNF-α) and CD68 in pulmonary tissues were analyzed quantitatively by immunohistochmistry and image analysis system. Results (1) The alveolitis and pulmonary fibrosis of rats in both SiO2 group and BLM group were became more serious gradually over time, HE staining under light microscope showed that BLM group on the 7th day had the most obvious alveolitis (2.814±0.832), the saturated picric acid sirius red staining under polarized light showed that BLM group on the 14th day had the worst pulmonary fibrosis (1284.57±554.72), which were significantly higher than those (103.69±18.29 and 111.78±37.45) in control group and SiO2 group on the 7th day (P＜0.05). (2) The results of immunohistochmistry examination indicated that the expression (17.100±1.831) of TNF-α in the BLM group on the 7th day was significantly higher than those (0.451 ±0.441, 7.909±1.275 and 13.506±1.454) in control group, SiO2 group on 7th day and BLM group on 14th day (P＜0.05). The expression (22.778 ±2.512) of TNF-a in the SiO2 group on the 14th day was significantly higher than those in control group, SiO2 group on 7th day and BLM group on 14th day (P＜0.05).The expression (134.941 ±35.951) of CD68 in the SiO2 group on the 14th day was significantly higher than those in control group,SiO2 group on 7th day and BLM group on 14th day(P＜0.05).Conclusion The early alveolitis of BLM-induced lung injury model was more serious than that of SiO2-induced lung injury model,and the fibrosis process of BLM-induced lung injury model was earlier than that of SiO2-induced lung injury model.TNF-α plays an important role in the conrse of both models.but macrophages is involved in Si06nduced pulmonary in a more continuous way than in BLM-induced pulmonary fibrosis.     

旋转脉动磁场对兔牙移动过程中牙周组织内血小板衍生生长因子A表达的影响

Objective To investigate the effects of rotating pulsed magnetic field on platelet derived growth factor a (PDGF-A) expression in periodontal tissue during tooth movement. Methods 30 white rabbits were random divided into 6 groups with 5 rabbits each group,including groups of 1,3,5,7,14 and 21 days. Under anesthesia condition by 2% pentobarbital sodium,the stainless coil springs were fixed between the first maxillary molar and the incisor producing the force of 80g. The experimental group was treated by rotating pulsed magnetic field and the force, the control group was only treated by t he force. The expression of PDGF-A was half-quantitatively investigated through immunohistochemical analysis. Results The expression of PDGF-AA in the experimental group enhanced apparently compared with that in the control group. There were significant differences among the 5,7, and 14day groups (5. 28 ± 0. 14 vs 2. 03 ±0. 18,7.63±0.27 vs 2. 84 ±0. 12,3.52 ±0. 16 vs 1.65 ±0.03;8.10±0.13 vs 4. 30 ±0. 21,13. 27 ±0. 31 vs 6.47 ± 0.15,5.66 ± 0.22 vs 3. 15 ± 0. 27, P ＜ 0. 05). The expression of PDGF-AA of 7 day group was higher than that of other groups. Conclusion Rotating pulsed magnetic field promotes the expression of PDGF-A in the periodontal tissue and alveolar bone remodeling.     

旋转脉动磁场对兔牙移动过程中牙周组织内血小板衍生生长因子A表达的影响

Objective To investigate the effects of rotating pulsed magnetic field on platelet derived growth factor a (PDGF-A) expression in periodontal tissue during tooth movement. Methods 30 white rabbits were random divided into 6 groups with 5 rabbits each group,including groups of 1,3,5,7,14 and 21 days. Under anesthesia condition by 2% pentobarbital sodium,the stainless coil springs were fixed between the first maxillary molar and the incisor producing the force of 80g. The experimental group was treated by rotating pulsed magnetic field and the force, the control group was only treated by t he force. The expression of PDGF-A was half-quantitatively investigated through immunohistochemical analysis. Results The expression of PDGF-AA in the experimental group enhanced apparently compared with that in the control group. There were significant differences among the 5,7, and 14day groups (5. 28 ± 0. 14 vs 2. 03 ±0. 18,7.63±0.27 vs 2. 84 ±0. 12,3.52 ±0. 16 vs 1.65 ±0.03;8.10±0.13 vs 4. 30 ±0. 21,13. 27 ±0. 31 vs 6.47 ± 0.15,5.66 ± 0.22 vs 3. 15 ± 0. 27, P ＜ 0. 05). The expression of PDGF-AA of 7 day group was higher than that of other groups. Conclusion Rotating pulsed magnetic field promotes the expression of PDGF-A in the periodontal tissue and alveolar bone remodeling.     

地塞米松预处理对大鼠光气吸入性肺损伤基质金属蛋白酶-9表达的影响

Objective To investigate the effects of dexamethasone on expression of matrix metalloproteinase-9 (MMP-9) in rats with acute lung injury induced by phosgene. Methods The rats were randomly divided into 3 groups: normal control group that consists of the rats with air exposure, phosgene group that consists of the rats with phosgene exposure and dexamethasone group that consists of the rats with phosgene exposure after 2.5 mg/kg dexamethasone being injected. Wet and dry ratio of the lung (W/D) was calculated, and leukocyte count and total protein content of bronchoalveolar lavage fluid (BALF) were recorded at 2 h after exposure. The concentrations of MMP-9 in the serum and BALF were measured by enzyme-linked immunosorbent assay. The pathologic changes of lung tissues were observed under light microscopy. The immunohistochemistry and the RT-PCR were used to detect the contents of MMP-9 in the lung tissue. Results Compared with phosgene group, the lung W/D, protein content and WBC count in of BALF dexamethasone group was significantly decreased (P＜0.01). MMP-9 levels of the serum and BALF in dexamethasone group were (4.799±0.043) μg/L and (15.052±0.029) μg/L, respectively, which were significantly lower than those [(9.439±0.100) and (20.640±0.446) μg/L] in phosgene group (P＜0.01). Compared with phosgene group (2.789±0.282), the expression level(1.183±0.260) of lung M MP-9 mRNA in dexamethasone group was significantly lower than that in phosgene group (P＜0.01).Histological experimental results showed the marked hyperemia and thickening of alveolar wallsand stroma cells infiltrating and more visible alveolar structure damage of alveolar walls in phosgene group while the alveolar structure and the alveolar walls were clear and slightly thickened with inflammatory cells in dexamethasone group. Immunohistochemical results showed that MMP-9 protein expression levels of lung and brochus tissues in normal control group and dexamethasone group were weakly positive, which in phosgene group were strongly positive. Conclusion Dexamethasone has a beneficial effects on acute lung injury induced by phosgene in rats due to the inhibiting MMP-9.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

细胞外信号调节蛋白激酶1/2信号通路对SiO2活化的肺成纤维细胞转化生长因子-β1表达的影响

目的 探讨细胞外信号调节蛋白激酶(ERK)信号通路对矽尘诱导的人胚肺成纤维细胞(HELF)中转化生长因子(TGF-β1)表达的影响及调控作用.方法 收集矽肺患者支气管肺泡灌洗液中的肺泡巨噬细胞(AM),在体外将AM分别用不含SiO2的DMEM培养液和含SiO2(50 μg/ml)的DMEM 培养液培养作用18 h后,将收集的AM条件培养上清液与HELF共同孵育.通过采用免疫细胞化学法检测应用ERK1/2抑制剂PD98059干预后SiO2活化的矽肺患者AM上清介导的HELF中TGF-β1表达的改变.结果 SiO2处理组HELF中TGF-β1的表达量(0.3227±0.0238)明显高于空白对照组(0.1637±0.0196)及AM对照组(0.2406±0.0225),而PD98059干预组TGF-β1的表达量(0.2711±0.0229)明显低于SiO2处理组,差异均有统计学意义(P＜0.05).结论 PD98059对由SiO2刺激的矽肺患者AM条件培养上清介导的HELF中TGF-β1的表达有一定抑制作用,ERK信号途径在一定程度上可能介导SiO2刺激的HELF细胞因子表达.

Abstract：

Objective To investigate the effect and regulation of extracellual signal-regulated kinase (ERK1/2)signaling pathway on the expression of transforming growth factor-β1 (TGF-β1) in human embryonic lung fibroblasts induced by SiO2. Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and in the presence or absence of SiO2 (50 ug/ml) exposition for 18h, and then the conditioned supernatants were used to incubate HELF. The expressions of TGF-β1 of the HELF acted with the conditioned AM supernatant fluid were detected with the immunocytochemistry method after treatment with PD98059 of inhibitor of ERK. Results The expression of TGF-β1 in HELF of the SiO2 treatment group(OD value is 0.322 7±0.023 8)exceed blank group (OD value is 0.163 7±0.019 6) and AM control group(OD value is 0.240 6±0.022 5) by the immunocytochemistry method. But the expression of TGF-β1 had reduction in some extent in the PD98059 intervention group (OD value is 0.271 l±0.022 9). The values were statistically different (P＜0.05). Conclusion ERK inhibitor PD98059 have inhibition effect on the expression of transforming growth factor-pi and expression of cytokine of human embryonic lung fibroblasts stimulated by SiO2, The study indicate that the proliferation and collagen prodction of HELF activated by SiO2 are mediated by ERK/MAPK signal pathway in some extent. PD98059 may antagonizes silica-induced lung fibrosis by inhibiting the expression of transforming growth factor-β1.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

The expression and meaning of TGF-β and TGIF in endometrosis

Objective To explore the role of TGF-βand TGIF in the pathogenesis of endometriosis. Methods The expression of TGF-β and TGIF was detected by immunohistochemistry method in the ectopic and eutopic endometrium of 30 cases with endometriosis (ec-topic endometrium group and eutopic endometrium group) and in the normal endometrium of 40 cases without endometriosis (control group). Result The expression of TGF -β in ectopic endometrium group was significantly higher than that in eutopic endometrium group and control group (P < 0.05). There was no significant difference in the expression of TGF- βbetween eutopic endometrium group and control group(P > 0.05). The expression of TGIF in ectopic endometrium group was significantly lower than that in eutopic endometrium group and control group( P <0.05). There was no significant difference in the expression of TGIF between eutopic endometrium group and control group(P > 0.05). There were negative correlation between the expressions of TGF - β and TGIF in ectopic endometrium group and eutopic endome-trium group(rs= - 0.769, - 0.549, P < 0.05). Conclusion The abnormal expression of TGF-β and TGIF in ectopic endometrium of pa-tients with endometriosis may be associated with the genesis and progression of endometriosis.